CN103768044B - Suppress the application of the reactive compound of DJ-1 dimerization - Google Patents
Suppress the application of the reactive compound of DJ-1 dimerization Download PDFInfo
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- 0 Oc(cccc1)c1N=*=CCC1=NC(c2ccccc2)=C(CC2=CC=CCC2)*1 Chemical compound Oc(cccc1)c1N=*=CCC1=NC(c2ccccc2)=C(CC2=CC=CCC2)*1 0.000 description 1
Abstract
The invention provides the application of one group of reactive compound in the medicine of preparation suppression DJ-1 dimerization.By finding one group of compound suppressing DJ-1 dimerization based on the area of computer aided virtual method of DJ-1 albumin crystal structure and technology, pharmaceutical chemistry knowledge and Pharmacological Activity Screening, totally 23.According to the compound that structural similarity coefficient T animoto value is greater than 0.85, there is similar biological activity principle, determine that organizing molecular structure of compounds similarity Tanimoto value with this is greater than 0.85, also there is similar suppression DJ-1 active.The present invention is with the DJ-1 albumen relevant to tumor development for object of study, and screening obtains having the compound suppressing DJ-1 protein dimerization and inhibition tumor cell propagation.These compounds not only can suppress the dimerization of DJ-1 albumen, and have stronger inhibit activities to tumor cell, have good drug development prospect.
Description
Technical field
The invention belongs to medical art, relate to one group and suppress the compound of DJ-1 protein dimerization and preparing the application in antitumor drug.
Background technology
Cancer, also known as malignant tumor, to rise in value the not normal and disease that causes of mechanism by controlling Growth of Cells.Cancerous cell except grow out of control except, also locally can invade surrounding normal tissue and even transfer to other parts of health via body-internal-circulation system or lymphsystem.Not only bring great misery to patient, and bring heavy economy and mental burden to family.
DJ-1 be by Japanese scholars Hiroyoshi in 1997 find a kind of albumen (DaisukeNagakubo, etal,
biochemicalandBiophysicalResearchCommunications, 1997,213,509-513).They are separated and obtain new cDNA from NIH3T3 cell, obtain DJ-1 by reverse transcription.It is one and forms dimer by 189 aminoacid.Each monomer secondary structure is by 8 α spirals, and 11 β-pleated sheets and 1 beta hairpin structure form.Dimer rely on 8 pairs of hydrogen bonds and a large amount of vanderWall ' s formed (K.Honbon,
journalofBiologicalChemistry, 2003,278 (33), 31380-31384).This albumen can promote the conversion of cell cycle, and relevant to H-Ras path, is identified as oncoprotein at first.In vivo, the protein binding such as DJ-1 needs and PIASx α and DJBP, maintenance activity (Q.Huai, etal,
fEBSletters, 2003,549,171-175).Research find, DJ-1 and oxidative stress in close relations.Wherein, Cys106 residue is the highest to the sensitivity stimulated, be active major site (T.Kinumi, etal,
biochemicalandBiophysicalResearchCommunications,2004,317 (3), 722-728).When in DJ-1 structure, L166p undergos mutation, the degraded of albumen can be caused, lose function, ROS be accumulated in a large number, causes relevant disease, such as, parkinson disease and Alzheimer (X, Tao, L, Tong.
journalofBiologicalChemistry, 2003,278 (33): 31372-31379).Recently, find the overexpression of DJ-1 and tumor occur relevant (H.Ren, etal,
cancerLetter, 2010,297 (1), 101-108).In ovarian cancer cell, detect DJ-1 high expressed, and the process LAN of DJ-1 makes the expression of Tumor growth inhibition factor phosphate and tensin congener (PTEN) to decline (B.Davidson, etal, HumanPathology, 2008,39 (1), 87-95); In breast cancer cell, DJ-1 not only suppresses the expression of PTEN, and promotes the phosphorylation of serine-threonine protein kinase enzyme (PKB/AKT) path.In addition, in other cancer cell such as lung carcinoma cell, all detect the high expressed of DJ-1.Nearest research shows, the high expressed of DJ-1, can reduce tumor cell to the sensitivity of chemotherapeutics (H.Ren, etal,
cancerLetter, 2010,297 (1), 101-108).
At present, the medicine being used for the treatment of cancer clinically much acts on the kinases relevant to tumor cell proliferation and albumen.Such as, paclitaxel (taxol), acts on tubulin; Gefitinib (gefitinib) and erlotinib (erlotinib), suppress the kinase activity relevant to tumor proliferation.But, because tumor cell is easily undergone mutation, produce drug resistance, cause the tumor cell of medicine to sudden change to lose activity.Generation and the development of DJ-1 and tumor have close association, but, at present still not for the compound or the related drugs report that suppress this protein active.Therefore, the antitumoral compounds developed for the novel structure of DJ-1 has great importance.
Along with the application of supercomputer in calculation biology and drug design that calculation function is powerful, the speed of SARS drug design and accuracy all obtain revolutionary raising, for Rational drug design provides effective technological means.Computer-Aided Drug Design based on biomacromolecule binding site or pharmacophore model, by carrying out virtual screening to compound database and find potential lead compound and carrying out biological activity test to it and compound structure optimization carrys out Development of Novel medicine.In Computer-Aided Drug Design, molecular simulation, molecular characterization and molecular conformation similarity system design and the protein-ligand interaction theoretical research based on molecular mutation and structure biology play crucial effect.In addition, the effect of virtual screening in the discovery and structure optimization of brand-new lead compound more and more comes into one's own, it is by the 3 d structure model of existing biomacromolecule or the Pharmacophore Model based on small molecule active part structure, the even more compound of in compound library up to a million is screened, choose the higher compound of marking as alternative lead compound, carry out the deeply exploitations such as bioactivity screening.Virtual screening considerably reduces the workload of compou nd synthesis and drug screening, time and fund input, improves efficiency and the accuracy of screening.Virtual screening can also complete the operation that some traditional methods cannot be carried out, as many condition and restrictive condition screening.Therefore, virtual screening obtains using more and more widely in new drug development research.
Summary of the invention
First object of the present invention is to provide the application of one group of compound in preparation DJ-1 dimerisation inhibitor.By finding one group of reactive compound suppressing DJ-1 dimerization based on the area of computer aided virtual method of DJ-1 albumin crystal structure and technology, pharmaceutical chemistry knowledge and Pharmacological Activity Screening.Particular compound following (being compound number of the present invention in bracket):
The chloro-5-Nitro-benzoic acid of 2--N '-(3-hydroxyl-naphthalene-2-carbonyl)-hydrazides (
1)
2-(4,5-diphenyl-1H-imidazoles-2-azo group)-phenol (
2)
5-(2-nitro-phenyl)-furan-2-carboxylic acid-(2,4-Dichloro-phenyl)-amide (
3)
4-hydroxyl-1-methyl-2-oxygen-1,2-dihydroquinoline-3-carboxylic acid N'-(the chloro-benzoyl of 2-)-hydrazides (
4)
(the bromo-benzoyl-amido of 4-)-acetic acid-2-(3-nitro-phenyl)-2-oxo-ethyl ester (
5)
The bromo-2-methyl of 4--2H-pyrazoles-3-carboxylic acid (5-chlorine-2-hydroxyl alkene benzyl)-hydrazides (
6)
3-hydroxyl naphthalene-2-carboxylic acid [1-(3-nitro-phenyl)-vinyl]-hydrazides (
7)
1-(2-Methyl-benzoyl)-3-(4-morpholine-4-methylphenyl)-thiourea (
8)
4-{ [(2-methyl-2,3-dioxy-1,3-dihydro-1H-iso-indoles-5-carbonyl)-amino]-methyl-benzoic acid (
9)
2-pyridin-4-yl-Cinchonic Acid-(3-methyl-thiophene-2-methylene)-hydrazides (
10)
2-pyridin-4-yl-Cinchonic Acid-pyridine-2-methylene-hydrazides (
11)
5-(4-nitro-1H-pyrazoles-1-methyl)-furan-2-formic acid (3-carbamoyl-4,5,6,7-tetrahydro benzo thiophene-2-base)-amide (
12)
2-BTA-1-base-acetamide-ethyl-thiourea (
13)
[6-oxygen-2-(N'-2-methylphenyl-guanidine radicals)-3,6-dihydro-pyrimidin-4-base]-methyl acetate (
14)
[5-amino-3-(4-chlorphenyl)-[1,2,4] triazol-1-yl]-furan-2-ketone (
15)
2-(6-Methoxy-benzothiazol-2-base-amino)-nicotinic acid (
16)
N-(the chloro-benzothiazole of 4--2-base-2-hydroxyl-nicotiamide (
17)
2-methoxyl group-N-[5-(3-nitro-phenyl)-[1,3,4] thiadiazoles-2-base]-Benzoylamide (
18)
5-bromine furan-2-carboxylic acid (between 2-benzo [1,3] dioxolen-5-base-1-carbamoyl vinyl)-amide (
19)
2-(the chloro-phenyl of 4--5-methyl isophthalic acid H-[1,2,4] triazole-3-carboxylic acid-(the fluoro-2-methylphenyl of 4-)-amide (
20)
5-methyl isophthalic acid-phenyl-1H-[1,2,3] triazole-4-carboxylic acid-(3-trifluoromethoxy-phenyl)-amide (
21)
N-{3-[(oxolane-2-methylene)-carbamoyl]-phenyl }-succinamic acid (
22)
1-(the fluoro-phenyl of 4--3-(2-trifluoromethyl-phenyl-[1,3]-dioxole-2-base)-urea (
23).
The compound being greater than 0.85 according to structural similarity coefficient T animoto value has similar bioactive principle (document M.A.Johnson, G.M.Maggiora, Eds.
conceptsandApplicationsofMolecularSimilarity; Wiley:NewYork, 1990
.y.C.Martin, etal.DoStructurallySimilarMoleculesHaveSimilarBiological Activity
journalofMedicinalChemistry, 2002,45,4350-4358.J.W.Godden, etal.
journalofChemicalInformationandComputerSciences2000,40,163-166 and X.-Q.Xie, etal.
journalofChemicalInformationandModeling2008,48,465-475), the present invention determines to organize with this compound that molecular structure of compounds similarity Tanimoto value is greater than 0.85, also has the activity of similar suppression DJ-1 dimerization.
Another object of the present invention is to provide the application of one group of reactive compound in the medicine of preparation suppression DJ-1 dimerization, and described reactive compound is:
The chloro-5-Nitro-benzoic acid of 2--N '-(3-hydroxyl-naphthalene-2-carbonyl)-hydrazides (
1)
The bromo-2-methyl of 4--2H-pyrazoles-3-carboxylic acid (5-chlorine-2-hydroxyl alkene benzyl)-hydrazides (
6)
3-hydroxyl naphthalene-2-carboxylic acid [1-(3-nitro-phenyl)-vinyl]-hydrazides (
7)
1-(2-Methyl-benzoyl)-3-(4-morpholine-4-methylphenyl)-thiourea (
8)
2-pyridin-4-yl-Cinchonic Acid-pyridine-2-methylene-hydrazides (
11)
5-(4-nitro-1H-pyrazoles-1-methyl)-furan-2-formic acid (3-carbamoyl-4,5,6,7-tetrahydro benzo thiophene-2-base)-amide (
12)
2-BTA-1-base-acetamide-ethyl-thiourea (
13)
[5-amino-3-(4-chlorphenyl)-[1,2,4] triazol-1-yl]-furan-2-ketone (
15)
2-methoxyl group-N-[5-(3-nitro-phenyl)-[1,3,4] thiadiazoles-2-base]-Benzoylamide (
18)
5-bromine furan-2-carboxylic acid (between 2-benzo [1,3] dioxolen-5-base-1-carbamoyl vinyl)-amide (
19)
2-(the chloro-phenyl of 4--5-methyl isophthalic acid H-[1,2,4] triazole-3-carboxylic acid-(the fluoro-2-methylphenyl of 4-)-amide (
20)
N-{3-[(oxolane-2-methylene)-carbamoyl]-phenyl }-succinamic acid (
22).
Found by the pharmacologically active experiment for tumor cell, described compound on tumor cell has good inhibit activities, wherein compound
1good inhibit activities is had to human cervical carcinoma Hela and human osteosarcoma KHOS tumor cell; Compound
8good inhibit activities is had to tumor cells such as human cervical carcinoma Hela cell's strain, human nasopharyngeal carcinoma KB cell strain, human osteosarcoma KHOS cell strain and human prostata cancer PC3 cell strains; Compound
11good inhibit activities is had to tumor cells such as human cervical carcinoma Hela cell's strain and human osteosarcoma KHOS cell strains; Compound
12good inhibit activities is had to tumor cells such as Human colorectal carcinoma HCT116 cell strain, human cervical carcinoma Hela cell's strain, human osteosarcoma KHOS cell strain, people's adenocarcinoma ovaries SKOV3 cell strain and human osteosarcoma U2OS cell strains; Compound
13good inhibit activities is had to tumor cells such as human cervical carcinoma Hela cell's strain and human osteosarcoma KHOS cell strains; Compound
20good inhibit activities is had to tumor cells such as human nasopharyngeal carcinoma KBKB cell strain and human osteosarcoma U2OS cell strains; Compound
6,
7,
15,
18,
19,
22good inhibitory action is had to tumor cell lines such as human pulmonary epithelial cells strain, Human colorectal carcinoma HCT116 cell strain, human cervical carcinoma Hela cell's strain, human hepatoma HepG2 cell's strain, human nasopharyngeal carcinoma KB cell strain, human osteosarcoma KHOS cell strain, human prostata cancer PC3 cell strain, people's adenocarcinoma ovaries SKOV3 cell strain and human osteosarcoma U2OS cell strains.Especially, compound
6to tumor cell people adenocarcinoma ovaries SKOV3 cell strain and compound
19to the inhibit activities IC of tumor cell Human colorectal carcinoma HCT116 cell strain
50value reaches 10 respectively
-6μ g/mL, compound
6to human cervical carcinoma Hela cell and compound
7to the inhibit activities IC of human prostata cancer PC3 cell
50value reaches 10 respectively
-3μ g/mL.
Same, according to the compound that structural similarity coefficient T animoto value is greater than 0.85, have similar bioactive principle, the present invention determines and the compound that above-mentioned 12 molecular structure of compounds similarity Tanimoto values are greater than 0.85, also has similar anti-tumor activity.
Another object of the present invention is to provide the method for above-mentioned 23 compounds of area of computer aided virtual screening, is achieved by the following scheme: utilize SybylX1.3 SARS drug design and simulation softward bag (SYBYL-X, version1.3; MolecularModelingSoftwarePackages, TriposAssociates, Inc., St.Louis, MO63144, USA, 2011), based on the albumin crystal structure (PDBID:1P5F of DJ-1, http://www.pdb.org), carry out drug design and virtual screening, wherein based on centered by protein D J-1 key amino acid Cys106 residue, around in 10 scopes, the close aminoacid of key amino acid and space comprising aminoacid Ala14, Glu15, Cys106, Arg48, Ala107, Thr125 and Leu128 etc. is selected to form potential binding pocket.
The present invention is when having no the research of bibliographical information DJ-1 inhibitor, first based on the three-dimensional crystalline structure of DJ-1, use the Method and Technology of the area of computer aided virtual screening of structure based, tested by pharmacologically active, found the suppression DJ-1 dimerization activity compound with good anti-tumor activity.The present invention with the DJ-1 albumen relevant to tumor development for object of study, use Method and Technology and the pharmaceutical chemistry relevant knowledge of Computer-Aided Drug Design, virtual screening obtains having the compound suppressing DJ-1 protein dimerization and inhibition tumor cell propagation.Preliminary pharmacological activity experiment shows: these compounds not only can suppress the dimerization of DJ-1 albumen, and have stronger inhibit activities to tumor cell, and is used for antitumor activity there are no reported in literature related compound.There is good drug development prospect.
Accompanying drawing explanation
Fig. 1 and Fig. 2 is the experimental result of Westernblotting, represents the experiment of 23 compounds suppression DJ-1 protein dimerization that virtual screening obtains."+DSS " expression adds protein-crosslinking agent two butanimide suberate; "-DSS " expression does not add two butanimide suberates; "-" represents blank.When adding DSS, if DJ-1 monomer does not have combined thing to degrade, again dimer can be formed; If the combined thing degraded of DJ-1 monomer, then again can not form the dimer of DJ-1, DJ-1 loses biological activity simultaneously.Fig. 1 and Fig. 2 display is compared with blank, and the dimerization of each compound to DJ-1 has obvious inhibit activities.
Detailed description of the invention
The present invention is further described in conjunction with the embodiments, but is not used for limiting the present invention.
embodiment 1
By literature survey, based on the albumin crystal structure (PDBID:1P5F, http://www.pdb.org) of DJ-1, to calculate figure Epidemiological Analysis and to infer the cavity being easy to ligand binding existed in DJ-1.Found by literature survey, Cys106 residue is most important for protein active.DJ-1 albumen is in close relations with the oxidative stress of body in vivo, and wherein Cys106 stimulates the most responsive to oxidation, is active major site.If Cys106 catalytic site can be occupied, just likely suppress the activity of DJ-1.Therefore, the present invention, centered by the Cys106 residue of albumen, finds potential binding pocket in 10 scopes around.Utilize Amber12 molecular dynamics simulation software, dynamics simulation is carried out to the potential binding pocket selected.Through optimization, heating, balance etc., the Molecular Dynamics Calculation of 30ns altogether, dynamics simulation and DJ-1, in conjunction with relevant, comprise the potential DJ-1 inhibitor binding pocket of key amino acid and the space close aminoacid composition of Ala14, Glu15, Cys106, Arg48, Ala107, Thr125 and Leu128 etc.Utilize SybylX1.3 software, and the result that binding kinetics calculates, simulate the binding pattern of potential DJ-1 active pocket.The pivotal role relevant to DJ-1 activity obtained based on calculating simulation and potential binding pocket, carry out the area of computer aided virtual screening of structure based to Specs compound database (http://www.specs, net/).
embodiment 2
Utilize SybylX1.3 SARS drug design and simulation softward bag (SYBYL-X, version1.3; MolecularModelingSoftwarePackages, TriposAssociates, Inc., St.Louis, MO63144, USA, 2011) FlexE-Dock and the Surflex-Dock software in, utilize molecular dynamics simulation to obtain to DJ-1 in conjunction with some relevant key amino acid effects and the binding pattern with lateral reactivity pocket, Specs compound library is carried out to the area of computer aided virtual screening of structure based.By the scoring system C-Score of Sybyl software self, marking rank is carried out to the docking capacity of each compound and DJ-1 in data base, choose rank front 10% compound.Then according to the result of docking of the physicochemical properties such as oil-water partition coefficient (ClogP<5), molecular weight (M.W.<500) of compound, compound and DJ-1, and in the structure diversity of compound and pharmaceutical chemistry, relevant knowledge selects compound.According to following formula (X.-Q.Xie, etal.
journalofChemicalInformationandModeling2008,48,465-475.) calculate the structural similarity coefficient T animoto value of two compounds:
Wherein
n a for the building stone that compound a has,
n b for the building stone that compound b has,
n ab for the building stone that compound a and b have jointly, the compound that classification compound likeness coefficient Tanimoto value is greater than 0.85.Meanwhile, the compound selecting not to be used for antitumor activity and drug development at bibliographical information is considered.Purchase virtual screening obtains the compound 100 that compound structure likeness coefficient Tanimoto value is less than 0.85, carries out pharmacologically active test.The reactive compound suppressing DJ-1 screening active ingredients to obtain is as follows:
The chloro-5-Nitro-benzoic acid of 2--N '-(3-hydroxyl-naphthalene-2-carbonyl)-hydrazides (
1)
2-(4,5-diphenyl-1H-imidazoles-2-azo group)-phenol (
2)
5-(2-nitro-phenyl)-furan-2-carboxylic acid-(2,4-Dichloro-phenyl)-amide (
3)
4-hydroxyl-1-methyl-2-oxygen-1,2-dihydroquinoline-3-carboxylic acid N'-(the chloro-benzoyl of 2-)-hydrazides (
4)
(the bromo-benzoyl-amido of 4-)-acetic acid-2-(3-nitro-phenyl)-2-oxo-ethyl ester (
5)
The bromo-2-methyl of 4--2H-pyrazoles-3-carboxylic acid (5-chlorine-2-hydroxyl alkene benzyl)-hydrazides (
6)
3-hydroxyl naphthalene-2-carboxylic acid [1-(3-nitro-phenyl)-vinyl]-hydrazides (
7)
1-(2-Methyl-benzoyl)-3-(4-morpholine-4-methylphenyl)-thiourea (
8)
4-{ [(2-methyl-2,3-dioxy-1,3-dihydro-1H-iso-indoles-5-carbonyl)-amino]-methyl-benzoic acid (
9)
2-pyridin-4-yl-Cinchonic Acid-(3-methyl-thiophene-2-methylene)-hydrazides (
10)
2-pyridin-4-yl-Cinchonic Acid-pyridine-2-methylene-hydrazides (
11)
5-(4-nitro-1H-pyrazoles-1-methyl)-furan-2-formic acid (3-carbamoyl-4,5,6,7-tetrahydro benzo thiophene-2-base)-amide (
12)
2-BTA-1-base-acetamide-ethyl-thiourea (
13)
[6-oxygen-2-(N'-2-methylphenyl-guanidine radicals)-3,6-dihydro-pyrimidin-4-base]-methyl acetate (
14)
[5-amino-3-(4-chlorphenyl)-[1,2,4] triazol-1-yl]-furan-2-ketone (
15)
2-(6-Methoxy-benzothiazol-2-base-amino)-nicotinic acid (
16)
N-(the chloro-benzothiazole of 4--2-base-2-hydroxyl-nicotiamide (
17)
2-methoxyl group-N-[5-(3-nitro-phenyl)-[1,3,4] thiadiazoles-2-base]-Benzoylamide (
18)
5-bromine furan-2-carboxylic acid (between 2-benzo [1,3] dioxolen-5-base-1-carbamoyl vinyl)-amide (
19)
2-(the chloro-phenyl of 4--5-methyl isophthalic acid H-[1,2,4] triazole-3-carboxylic acid-(the fluoro-2-methylphenyl of 4-)-amide (
20)
5-methyl isophthalic acid-phenyl-1H-[1,2,3] triazole-4-carboxylic acid-(3-trifluoromethoxy-phenyl)-amide (
21)
N-{3-[(oxolane-2-methylene)-carbamoyl]-phenyl }-succinamic acid (
22)
1-(the fluoro-phenyl of 4--3-(2-trifluoromethyl-phenyl-[1,3]-dioxole-2-base)-urea (
23).
embodiment 3compound suppresses the experiment of DJ-1 dimerization:
Test antibody used purchased from Santa company (SantaCruz, CA, USA) and Cellsignaling company (CellsignalingTechnology); The IgG of horseradish peroxidase-labeled goat-anti rabbit and the IgG of sheep anti mouse is purchased from Calbiochem company (Darmstadt, Germany); ECL test kit is purchased from Pierce company (Rockford, IL, USA); ECLplus reagent chromogenic agents box is purchased from AmershamBiosciences company (ArlingtonHeights, IL, USA).
Employing Westernblotting method measures the effect that 23 compounds screened suppress DJ-1 dimerizations:
Use RIPAbuffer(50mMTris-HClpH7.4,150mMNaCl, 1mMEDTA, 25mM β-phosphoglycerol, 1mMPMSF, 0.1mM vanadic acid sodium, 5 μ g/mlleupeptin, 1%NP40,1%TritonX-100,0.2%SDS) DJ-1 purifying protein (4mg/mL) is doubly diluted by 1:100,10 μ l/EP pipe subpackages, add each compound that virtual screening obtains respectively, make final compound concentration be 10 μMs.After hatching 2 hours on ice, add the intramolecular crosslinking agent two butanimide suberate (disuccinimidylsuberate, DSS) of 100 μMs, hatch 1h, 15mMTris-HCl(pH7.5 for 4 DEG C) 15min cessation reaction.Add 5 μ lloadingbuffer(5 ×), boil 5min and make albuminous degeneration, Westernblotting detects DJ-1 dimer formational situation (see accompanying drawing 1 and accompanying drawing 2).
embodiment 4the inhibit activities test of compound on tumor cell:
Choose human pulmonary epithelial cells strain, Human colorectal carcinoma HCT116 cell strain, human cervical carcinoma Hela cell's strain, human hepatoma HepG2 cell's strain, human nasopharyngeal carcinoma KB cell strain, human osteosarcoma KHOS cell strain, human prostata cancer PC3 cell strain, people's adenocarcinoma ovaries SKOV3 cell strain and human osteosarcoma U2OS cell strain, carry out inhibiting tumour cells active testing.
The present invention's tumor cell line used, comprise the tumor cell lines such as human pulmonary epithelial cells strain, Human colorectal carcinoma HCT116 cell strain, human cervical carcinoma Hela cell's strain, human hepatoma HepG2 cell's strain, human nasopharyngeal carcinoma KB cell strain, human osteosarcoma KHOS cell strain, human prostata cancer PC3 cell strain, people's adenocarcinoma ovaries SKOV3 cell strain and human osteosarcoma U2OS cell strain, all buy in Shanghai cell institute; Cell culture Tissue Culture Dish used, Tissue Culture Plate are all purchased from Corning-Costar company (NewYork, USA); Sulforhodamine B(SulforhodamineB, SRB) and dimethyl sulfoxide (DMSO) available from Sigma; Dulbecco ' sModifiedEagleMedium(DMEM) powder is purchased from GibcoBRL company (LifeTechnologies, GrandIsland, NY, USA); Hyclone (FetalBovineSerum, FBS) is purchased from Hangzhou Ilex purpurea Hassk.[I.chinensis Sims company and GibcoBRL company (LifeTechnologies, GrandIsland, NY, USA).
Experimental implementation of the present invention is for human cervical carcinoma cell Hela, and all compounds of Simultaneously test are to the inhibit activities of above-mentioned each tumor cell.Experiment use tumor cell line, be incubated at DMEM culture fluid, containing the hyclone of 10% deactivation, penicillin 100IU/ml and streptomycin 100
/ ml, in 37 DEG C of (5%CO
2, 95% air), cultivate under saturated humidity environment.
Sulfonyl rhodamine-B assay (srb assay) measures the inhibitory action of compound on tumor cell proliferation:
To take the logarithm the tumor cell line of trophophase, be inoculated in 96 well culture plates by 4000/ porocyte, negative control group (DMSO) and compound treatment group are set.Compound maximum concentration is 50 μ g/ml, 5 times of gradient dilutions, altogether 5 concentration, the multiple hole of each concentration three.Compound effects cell is after 72 hours, discard culture fluid, every hole adds 10% trichloroacetic acid (TCA) the solution 100 μ l fixed cell of pre-cooling, 4 DEG C of refrigerators place 1 hour, the each hole of culture plate is with deionized water wash 5 times, to remove trichloroacetic acid solution, in atmosphere after drying, every hole adds SRB solution (4mg/ml) the 50 μ l of 1% peracetic acid formulation, ambient temperatare puts 20 minutes, discard in each hole and wash 5 times with 1% acetic acid after liquid, air drying after clean unconjugated SRB dyestuff, every hole adds the 10mMTris-base(tri methylol amino methane of pH=10.5) solution 100 μ l dissolves, oscillator plate vibrates 5 minutes, absorbance OD value is measured under microplate reader 515nm wavelength.
Part of compounds is to the IC of above-mentioned each tumor cell (human pulmonary epithelial cells strain, Human colorectal carcinoma HCT116 cell strain, human cervical carcinoma Hela cell's strain, human hepatoma HepG2 cell's strain, human nasopharyngeal carcinoma KB cell strain, human osteosarcoma KHOS cell strain, human prostata cancer PC3 cell strain, people's adenocarcinoma ovaries SKOV3 cell strain and human osteosarcoma U2OS cell strain)
50(μ g/mL) value is in table 1.
Find out from accompanying drawing 1 and table 1: (1) all compounds can suppress the dimerization of DJ-1.(2) great majority screen the compound on tumor cell obtained good inhibit activities.Wherein, compound
1good inhibit activities is had to Hela and KHOS tumor cell line; Compound
8good inhibit activities is had to tumor cell lines such as Hela, KB, KHOS and PC3; Compound 11 pairs of Hela and KHOS tumor cell lines have good inhibit activities; Compound
12good inhibit activities is had to tumor cell lines such as HCT116, Hela, KHOS, SKOV3 and U2OS; Compound
13good inhibit activities is had to Hela and KHOS cell strain; Compound
20good inhibit activities is had to KB and U2OS cell strain; (4) compound
6,
7,
15,
18,
19,
22good inhibitory action is had to all tumor cells.Especially, compound
6to tumor cell SKOV3 and compound
19to the inhibit activities IC of tumor cell HCT116
50value reaches 10 respectively
-6μ g/mL, compound
6to tumor cell Hela and compound
7to the inhibit activities IC of tumor cell PC3
50value reaches 10 respectively
-3μ g/mL.(5) generally speaking, the compound that virtual screening obtains has good antitumor application prospect, thus has good potential commercial value.(6) according to the compound that structural similarity coefficient T animoto value is greater than 0.85, there is similar biological activity principle (document M.A.Johnson, G.M.Maggiora, Eds.
conceptsandApplicationsofMolecularSimilarity; Wiley:NewYork, 1990
.y.C.Martin, etal.DoStructurallySimilarMoleculesHaveSimilarBiological Activity
journalofMedicinalChemistry, 2002,45,4350-4358.J.W.Godden, etal.
journalofChemicalInformationandComputerSciences2000,40,163-166 and X.-Q.Xie, etal.
journalofChemicalInformationandModeling2008,48,465-475), the compound being greater than 0.85 with above-claimed cpd structural similarity coefficient T animoto value, has similar anti-tumor activity.
Without the need to further elaboration, believe content disclosed before employing, those skilled in the art can apply the present invention to greatest extent.Therefore, embodiment is above interpreted as only illustrating, but not limits the scope of the invention by any way.
Claims (3)
1. one group of compound application in preparation DJ-1 dimerisation inhibitor, particular compound is as follows:
The chloro-5-Nitro-benzoic acid of 2--N '-(3-hydroxyl-naphthalene-2-carbonyl)-hydrazides (
1)
2-(4,5-diphenyl-1H-imidazoles-2-azo group)-phenol (
2)
5-(2-nitro-phenyl)-furan-2-carboxylic acid-(2,4-Dichloro-phenyl)-amide (
3)
4-hydroxyl-1-methyl-2-oxygen-1,2-dihydroquinoline-3-carboxylic acid N'-(the chloro-benzoyl of 2-)-hydrazides (
4)
(the bromo-benzoyl-amido of 4-)-acetic acid-2-(3-nitro-phenyl)-2-oxo-ethyl ester (
5)
The bromo-2-methyl of 4--2H-pyrazoles-3-carboxylic acid (5-chlorine-2-hydroxyl alkene benzyl)-hydrazides (
6)
3-hydroxyl naphthalene-2-carboxylic acid [1-(3-nitro-phenyl)-vinyl]-hydrazides (
7)
1-(2-Methyl-benzoyl)-3-(4-morpholine-4-methylphenyl)-thiourea (
8)
4-{ [(2-methyl-2,3-dioxy-1,3-dihydro-1H-iso-indoles-5-carbonyl)-amino]-methyl-benzoic acid (
9)
2-pyridin-4-yl-Cinchonic Acid-(3-methyl-thiophene-2-methylene)-hydrazides (
10)
2-pyridin-4-yl-Cinchonic Acid-pyridine-2-methylene-hydrazides (
11)
5-(4-nitro-1H-pyrazoles-1-methyl)-furan-2-formic acid (3-carbamoyl-4,5,6,7-tetrahydro benzo thiophene-2-base)-amide (
12)
2-BTA-1-base-acetamide-ethyl-thiourea (
13)
[6-oxygen-2-(N'-2-methylphenyl-guanidine radicals)-3,6-dihydro-pyrimidin-4-base]-methyl acetate (
14)
[5-amino-3-(4-chlorphenyl)-[1,2,4] triazol-1-yl]-furan-2-ketone (
15)
2-(6-Methoxy-benzothiazol-2-base-amino)-nicotinic acid (
16)
N-(the chloro-benzothiazole of 4--2-base)-2-hydroxyl-nicotiamide (
17)
2-methoxyl group-N-[5-(3-nitro-phenyl)-[1,3,4] thiadiazoles-2-base]-Benzoylamide (
18)
5-bromine furan-2-carboxylic acid (between 2-benzo [1,3] dioxolen-5-base-1-carbamoyl vinyl)-amide (
19)
2-(the chloro-phenyl of 4-)-5-methyl isophthalic acid H-[1,2,4] triazole-3-carboxylic acid-(the fluoro-2-methylphenyl of 4-)-amide (
20)
5-methyl isophthalic acid-phenyl-1H-[1,2,3] triazole-4-carboxylic acid-(3-trifluoromethoxy-phenyl)-amide (
21)
N-{3-[(oxolane-2-methylene)-carbamoyl]-phenyl }-succinamic acid (
22)
1-(the fluoro-phenyl of 4-)-3-(2-trifluoromethyl-phenyl-[1,3]-dioxole-2-base)-urea (
23).
2. one group is suppressed the reactive compound of DJ-1 dimerization preparing the application in antitumor drug, and particular compound is as follows:
The chloro-5-Nitro-benzoic acid of 2--N '-(3-hydroxyl-naphthalene-2-carbonyl)-hydrazides (
1)
The bromo-2-methyl of 4--2H-pyrazoles-3-carboxylic acid (5-chlorine-2-hydroxyl alkene benzyl)-hydrazides (
6)
3-hydroxyl naphthalene-2-carboxylic acid [1-(3-nitro-phenyl)-vinyl]-hydrazides (
7)
1-(2-Methyl-benzoyl)-3-(4-morpholine-4-methylphenyl)-thiourea (
8)
2-pyridin-4-yl-Cinchonic Acid-pyridine-2-methylene-hydrazides (
11)
5-(4-nitro-1H-pyrazoles-1-methyl)-furan-2-formic acid (3-carbamoyl-4,5,6,7-tetrahydro benzo thiophene-2-base)-amide (
12)
2-BTA-1-base-acetamide-ethyl-thiourea (
13)
[5-amino-3-(4-chlorphenyl)-[1,2,4] triazol-1-yl]-furan-2-ketone (
15)
2-methoxyl group-N-[5-(3-nitro-phenyl)-[1,3,4] thiadiazoles-2-base]-Benzoylamide (
18)
5-bromine furan-2-carboxylic acid (between 2-benzo [1,3] dioxolen-5-base-1-carbamoyl vinyl)-amide (
19)
2-(the chloro-phenyl of 4-)-5-methyl isophthalic acid H-[1,2,4] triazole-3-carboxylic acid-(the fluoro-2-methylphenyl of 4-)-amide (
20)
N-{3-[(oxolane-2-methylene)-carbamoyl]-phenyl }-succinamic acid (
22);
Wherein, compound
1anti-human cervical cancer Hela cells tumor cell line and human osteosarcoma KHOS tumor cell line; Compound
8anti-human s strain, human nasopharyngeal carcinoma KB cell strain, human osteosarcoma KHOS cell strain and human prostata cancer PC3 cell strain; Compound
11anti-human s strain and human osteosarcoma KHOS cell strain; Compound
12anti-human colorectal cancer HCT116 cell strain, human cervical carcinoma Hela cell's strain, human osteosarcoma KHOS cell strain, people's adenocarcinoma ovaries SKOV3 cell strain and human osteosarcoma U2OS cell strain; Compound
13the tumor cells such as anti-human s strain and human osteosarcoma KHOS cell strain; Compound
20anti-human nasopharyngeal carcinoma KB cell strain and human osteosarcoma U2OS cell strain; Compound
6,
7,
15,
18,
19,
22anti-human lung adenocarcinoma A549 cell line, Human colorectal carcinoma HCT116 cell strain, human cervical carcinoma Hela cell's strain, human hepatoma HepG2 cell's strain, human nasopharyngeal carcinoma KB cell strain, human osteosarcoma KHOS cell strain, human prostata cancer PC3 cell strain, people's adenocarcinoma ovaries SKOV3 cell strain and human osteosarcoma U2OS cell strain.
3. the application of one group of compound according to claim 1 in preparation DJ-1 dimerisation inhibitor, it is characterized in that, the screening technique of described 23 compounds, realized by following scheme: utilize SybylX1.3 SARS drug design and simulation softward bag, based in the albumin crystal structure of DJ-1, centered by key amino acid Cys106 residue, around in 10 scopes, comprise aminoacid Ala14, Glu15, Cys106, Arg48, Ala107, Thr125, potential binding pocket is formed at the close aminoacid of interior key amino acid and space with Leu128, carry out drug design and virtual screening, obtain the reactive compound suppressing DJ-1 dimerization.
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| CN109700794B (en) * | 2017-03-01 | 2021-01-08 | 浙江大学 | Hydrazone structure type androgen receptor antagonist and application thereof |
| CN108329232B (en) * | 2018-01-08 | 2021-04-16 | 浙江大学 | Hydrazide derivative and application thereof |
| CN113929591A (en) * | 2021-10-09 | 2022-01-14 | 中国科学技术大学 | Inhibitors with antiproliferative activity |
| CN115340535B (en) * | 2022-07-28 | 2024-03-08 | 广东省生殖科学研究所(广东省生殖医院) | Compounds with DJ-1 protein stabilizer effect and application thereof |
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| CN101433545A (en) * | 2008-12-23 | 2009-05-20 | 北京大学 | Use of bioflavanoid or polyphenolic substance for treating parkinson's disease |
| CN102134245A (en) * | 2009-12-24 | 2011-07-27 | 沈阳药科大学 | Tetralin isoquinoline compounds as well as preparation methods and applications thereof |
| KR20120060051A (en) * | 2010-12-01 | 2012-06-11 | 대구대학교 산학협력단 | Composition and Kit for Detecting Biomarkers for Obesity in Adipose Tissue |
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| WO2006104749A2 (en) * | 2005-03-25 | 2006-10-05 | Caritas St. Elizabeth Medical Center Of Boston, Inc. | Compositions and methods for detecting compounds that treat a neurological disorder |
| CN101433545A (en) * | 2008-12-23 | 2009-05-20 | 北京大学 | Use of bioflavanoid or polyphenolic substance for treating parkinson's disease |
| CN102134245A (en) * | 2009-12-24 | 2011-07-27 | 沈阳药科大学 | Tetralin isoquinoline compounds as well as preparation methods and applications thereof |
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